Preparation and applications of Doxorubicin-loading active targeting albumin nanometer carrier
A technology of albumin nanoparticles and albumin nanoparticles, which is applied in the field of anti-tumor targeted drug research to achieve the effects of ensuring activity, reducing immunogenicity, and improving targeting
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Embodiment 1
[0032] (1) Prepare doxorubicin-loaded nanoparticles: dissolve 40 mg of albumin in 2 ml of water and pass through a 0.22 μm filter; add 800 μl of 0.5% doxorubicin hydrochloride solution, then add 0.05 mol / LpH9 sodium carbonate-sodium bicarbonate Buffer 1.2ml, stirred at room temperature for 2h. Then 16ml of absolute ethanol was added dropwise at a speed of 0.5ml / min with an infusion set, and 23μl of 25% glutaraldehyde solution was added to solidify for 16h. After solidification, the reaction solution was vacuum rotary evaporated at 45°C to remove ethanol, then ultrasonically dispersed with distilled water, centrifuged at 1,2000 rpm for 15 min, and the supernatant was discarded after centrifugation. Wash the centrifuged precipitate three times in the same way, and ultrasonically disperse with the same volume of distilled water each time. The obtained doxorubicin-loaded albumin nanoparticles (DOX-NPs) had an average particle size of 160.1 nm and a polydispersity coefficient of a...
Embodiment 2
[0035] (1) Add 300μl 250mM borax / 250mM sodium chloride and 300μl 100mM dithiothreitol (pH8.0) to 2.4ml cetuximab (5mg / ml), incubate at 37°C for 45min, desalt by ultrafiltration and centrifugation, and purify the reduced Antibody. Adjust the antibody concentration (2mg / ml), take 4ml, add MC-Val-Cit-PAB-DOX / DMF (5mg / ml) 0.6ml, let it stand on ice for 1h, add 490μl cysteine (10mg / ml, containing Diethylenetriaminepentaacetic acid / PBS), and then rest for 45min. Desalting by ultrafiltration and centrifugation. Obtain the drug-antibody conjugate cetuximab-dipeptide doxorubicin (Cetuximab-vcDOX), measure the absorbance A280 and A495 of the drug-antibody conjugate at 280nm and 495nm by UV spectrophotometry, and pass the doxorubicin The standard curve at 280nm and 495nm of cetuximab and cetuximab calculates that the molar ratio of doxorubicin / cetuximab is 5.5, that is, 5.5 doxorubicin is connected to each cetuximab;
[0036] (2) Take 0.5 ml of 1% albumin solution, and slowly add 1....
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